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Tumor suppressor protein p53 exerts negative transcriptional regulation on human sodium iodide symporter gene expression in breast cancer

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Abstract

Purpose

Aberrant expression of human sodium iodide symporter (NIS) in breast cancer (BC) is well documented but the transcription factors (TF) regulating its aberrant expression is poorly known. We identify the presence of three p53 binding sites on the human NIS promoter sequence by conducting genome-wide TF analysis, and further investigate their regulatory role.

Methods

The differences in transcription and translation were measured by real-time PCR, luciferase reporter assay, site-directed mutagenesis, in vivo optical imaging, and chromatin immunoprecipitation. The relation of NIS and p53 in clinical samples was judged by TCGA data analysis and immunohistochemistry.

Results

Overexpression of wild-type p53 as a transgene or pharmacological activation by doxorubicin drug treatment shows significant suppression of NIS transcription in multiple BC cell types which also results in lowered NIS protein content and cellular iodide intake. NIS repression by activated p53 is further confirmed by non-invasive bioluminescence imaging in live cell and orthotropic tumor model. Abrogation of p53-binding sites by directional mutagenesis confirms reversal of transcriptional activity in wild-type p53-positive BC cells. We also observe direct binding of p53 to these sites on the human NIS promoter. Importantly, TCGA data analysis of NIS and p53 co-expression registers an inverse relationship between the two candidates.

Conclusion

Our data for the first time highlight the role of p53 as a negative regulator of functional NIS expression in BC, where the latter is a potential targeted radioiodine therapy candidate. Thus, the study provides an important insight into prospective clinical application of this approach that may significantly impact the patient with mutant versus wild-type p53 profile.

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Abbreviations

atRA:

All-trans retinoic acid

BC:

Breast cancer

ChIP:

Chromatin immunoprecipitation

Dox:

Doxorubicin

IHC:

Immunohistochemistry

NIS:

Sodium iodide symporter

NUE:

NIS upstream enhancer

RAR:

Retinoic acid receptor

RXR:

Retinoid X receptor

TNBC:

Triple-negative breast cancer

TF:

Transcription factor

TSH:

Thyroid-stimulating hormone

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Acknowledgements

We acknowledge Dr. Kenneth Ain, USA, Dr. Sorab Dalal, India for providing various plasmids used in the study. We acknowledge the prior intramural research funding from TMC Woman Cancer Initiative (WCI) (#82) to AD, as well as various institutional facilities supporting this work.

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Authors and Affiliations

  1. Molecular Functional Imaging Lab, Tata Memorial Centre, ACTREC, Sector 22, Kharghar, Navi Mumbai, 410210, India

    Madhura G. Kelkar, Abhishek Derle, Sushmita Chatterjee & Abhijit De

  2. Imaging Cell Signaling and Therapeutics Lab, Tata Memorial Centre, ACTREC, Navi Mumbai, India

    Bhushan Thakur & Pritha Ray

  3. Homi Bhabha National Institute, Anushakti Nagar, Mumbai, India

    Madhura G. Kelkar, Bhushan Thakur, Pritha Ray & Abhijit De

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  1. Madhura G. Kelkar
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  2. Bhushan Thakur
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Correspondence to Abhijit De.

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Kelkar, M.G., Thakur, B., Derle, A. et al. Tumor suppressor protein p53 exerts negative transcriptional regulation on human sodium iodide symporter gene expression in breast cancer. Breast Cancer Res Treat 164, 603–615 (2017). https://doi.org/10.1007/s10549-017-4297-2

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